Population density does not affect seasonal regulation of reproductive physiology in male water voles.

Most small rodent species display cyclic fluctuations in their population density. The mechanisms behind these cyclical variations are not yet clearly understood. Density-dependent effects on reproductive function could affect these population variations. The fossorial water vole ecotype, Arvicola terrestris, exhibits multi-year cyclical dynamics with outbreak peaks. Here, we monitored different water vole populations over 3 years, in spring and autumn, to evaluate whether population density is related to male reproductive physiology. Our results show an effect of season and inter-annual factors on testis mass, plasmatic testosterone level, and androgen-dependent seminal vesicle mass. By contrast, population density does not affect any of these parameters, suggesting a lack of modulation of population dynamics by population density.

Prenatal programming by testosterone of follicular theca cell functions in ovary.

In mammalian ovaries, the theca layers of growing follicles are critical for maintaining their structural integrity and supporting androgen synthesis. Through combining the postnatal monitoring of ovaries by abdominal magnetic resonance imaging, endocrine profiling, hormonal analysis of the follicular fluid of growing follicles, and transcriptomic analysis of follicular theca cells, we provide evidence that the exposure of ovine fetuses to testosterone excess activates postnatal follicular growth and strongly affects the functions of follicular theca in adulthood. Prenatal exposure to testosterone impaired androgen synthesis in the small antral follicles of adults and affected the expression in their theca cells of a wide array of genes encoding extracellular matrix components, their membrane receptors, and signaling pathways. Most expression changes were uncorrelated with the concentrations of gonadotropins, steroids, and anti-Müllerian hormone in the recent hormonal environment of theca cells, suggesting that these changes rather result from the long-term developmental effects of testosterone on theca cell precursors in fetal ovaries. Disruptions of the extracellular matrix structure and signaling in the follicular theca and ovarian cortex can explain the acceleration of follicle growth through altering the stiffness of ovarian tissue. We propose that these mechanisms participate in the etiology of the polycystic ovarian syndrome, a major reproductive pathology in woman.

Facial expression and oxytocin as possible markers of positive emotions in horses.

Behavioural and physiological markers of discrete positive emotions remain little investigated in animals. To characterise new markers in horses, we used tactile stimulations to induce emotional situation of contrasting valence. In the Gentle grooming group (G, N = 13) horses were gently groomed during 11 sessions on the body areas they appreciated the most. Horses in the Standard grooming group (S, N = 14) were groomed using a fixed procedure, reported to induce avoidance reactions in some horses. At session 11, G horses expressed significantly more contact-seeking behaviours than S horses, which expressed significantly more avoidance behaviours. This result suggests positive emotions in G horses and negative emotions in S horses. Blood cortisol, oxytocin, heart rate and heart rate variability never differed between before and after the grooming session. However, after the 11 sessions, basal oxytocin levels were lower in the G than in the S group. This difference was unexpected, but supports studies showing that a low level of basal oxytocin could be a marker of better well-being. Analyses of facial expressions during grooming revealed significant differences between groups. These expressions appear to be more sensitive than behavioural indicators because they alone enabled differentiating emotions according to the group when horses were re-exposed to neutral grooming one year after the treatment.

Reproductive parameters of donkey jacks undergoing puberty.

In male donkeys, puberty and the related events have been poorly characterized. The aim of this study was to evaluate the age at which male donkeys reach puberty, and characterize age associated changes in testicular size, testicular blood flow, serum testosterone concentration and semen quality. Every two months, starting at 6 months and finishing at 24 months of age, five male donkeys born in May to July were subjected to B-mode ultrasound examination to assess testicular size and scrotum content and blood serum sampling for testosterone concentration. From the age of 8 months, pulsed Doppler was employed to evaluate blood flow in the testicular artery. Testosterone serum concentration was evaluated via RIA. From the age of 12 months, monthly semen collections were attempted and semen was evaluated for sperm number, motility and morphology. Onset of puberty was defined as the first ejaculate containing ≥50 × 106 spermatozoa with ≥10% total motility. One of the donkeys was excluded from the statistical analyses due to a hydrocele presented during the study. Testes width was affected by age (P < 0.0001) and after an initial plateau increased linearly from 10 months of age. Pulsatility and resistivity indexes were also affected by age (P < 0.01), being significantly higher at 14 months than at 24 months. Testosterone serum concentration was affected by age (P < 0.0001) and was significantly lower at 6 months (0.1 ng/ml) compared to 22-24 months (≥0.8 ng/ml). Spermatozoa appeared in the ejaculate at a mean age of 18.7 months and puberty was attained between 19 and 20 months of age (mean: 19.5 months), between January and February. In conclusion, late spring born Amiata donkey colts reached puberty at 19-20 months of age. Puberty was accompanied by changes in testicular size, testicular blood flow and serum testosterone concentration.

All bonds are not alike: A psychoendocrine evaluation of infant attachment.

Characteristics of attachment were assessed in peer- and object-reared lambs, and compared to mothered subjects by taking into consideration distress, proximity seeking, and exploration during two separation-reunion tests in both the familiar and a novel environment. Plasma cortisol and oxytocin were assayed as physiological indicators of stress and being comforted during the separation-reunion test. Rewarding properties of the familiar figures were also determined in a conditioned place preference-like paradigm. Between-group analysis revealed the existence of secure attachment with the mother, alteration of secure attachment with the peer and weaker attachment with the object. Weaker attachment was expressed by a lack of distress during separation in the home pen and no preference for the place conditioned with the familiar object. Elevated basal plasma oxytocin levels, but not cortisol, observed in maternally deprived lambs were more likely linked to the absence of a maternal figure rather than social comfort during reunion.

Physiological changes in the peri-partum period and colostral IgG transfer in prolific D'man sheep: effects of parity and litter size.

The aim of this work was to assess maternal and neonatal changes in plasma proteins, glucose and cortisol and to quantify the colostral immunoglobulin G (IgG) transfer in the peri-partum period in D'man sheep, a prolific breed, taking into account the parity of the ewe. The concentrations of proteins and glucose were high in the ewes on day 7 and at lambing before decreasing. Likewise, cortisol plasma concentration was maximal during the 6 h following lambing and dropped at 12 h. Protein and glucose concentrations were low in lambs at 1 h of birth after which they increased. By contrast, cortisol level was the highest during the first 12 h of birth and then decreased. The colostral IgG level was high at lambing and dropped by over 87 % from 1 to 48 h post-partum. In the newborn, the plasma IgG concentration was lowest at birth and increased rapidly during the first 24 h of birth. Parity influenced maternal physiology with multiparous ewes having the lowest concentrations of proteins, glucose, IgG and cortisol, but the highest colostrum IgG level. Accordingly, lambs born from primiparous ewes had lower protein, glucose and plasma IgG concentrations than lambs born from multiparous ewes. The main outcome of this study was that lambs born from primiparous ewes are characterized by the lowest physiological indices and this may influence their survival chance.

Multiple exposures to familiar conspecific withdrawal is a novel robust stress paradigm in ewes.

Paradigms used so far to study the effects of social isolation in sheep confound the effects of social isolation with those of other stressors (e.g. new environment) and showed contradictory effects after multiple social isolations. We propose here to characterize and examine the repeatable effects of social isolation induced by the familiar conspecific withdrawal (FCW). This latter test consists of socially isolating the ewe by the removal of group mates from the room test for 3 h. Behavioral and endocrine responses of adult ovariectomized-estradiol implanted ewes were compared 90 min before and 90 min after FCW, which was applied three times every fourteen days. We observed that each FCW induced significant increases in plasma cortisol level, in the number of vocalizations, foot pawing, circling attempts and a significant decrease in time spent lying down. An increase in plasma cortisol levels and decrease in duration of maintenance behaviors were significantly lower after the third FCW than after the first one. These differences could be explained by higher plasma cortisol levels and lower duration of maintenance behaviors before the third FCW than before the first FCW suggesting an anticipation of the social isolation period. These data indicate that social isolation is sufficient to induce distress with stable stressful responses after multiple exposures to familiar conspecific withdrawal.

Anti-Mullerian hormone as a predictive endocrine marker for embryo production in the goat.

Recently, we demonstrated the relationship between anti-Müllerian hormone (AMH) circulating concentrations, ovarian follicles, and embryo production in cattle. However, they have not yet been established in a species with a seasonal breeding activity. Thus, goats were subjected to repeated in vivo embryo production during the breeding season, at the end of the breeding season, and at the end of the anestrus season. Embryo production after FSH treatment was highly repeatable for each goat. Plasma AMH concentrations, measured before the first FSH treatment, were highly correlated with the number of collected, transferable, and freezable embryos, resulting from the three sessions of embryo production. Plasma AMH concentrations transiently decreased after each exogenous FSH treatment, but they showed little change with season, and no relationship was observed between AMH and endogenous FSH concentrations during seasonal transitions. Follicles of 1-5 mm in diameter were the main target of the FSH treatment and were major contributors to circulating AMH concentrations. Granulosa cell AMH expression decreased as the follicle approached terminal development, while the expression of maturation markers (CYP19A1 and FSHR) increased. In conclusion, circulating AMH concentrations can be predictive of the capacity of a donor goat to produce high or low numbers of high-quality embryos. This prediction could be accurately made from a single blood measurement of AMH during either breeding or anestrus seasons. Variability in the number of gonadotropin-responsive follicles of 1-5 mm in diameter between individuals resulted in the differences in circulating AMH concentrations measured between individuals.

Differential protein expression assessed by two-dimensional gel electrophoresis for two wheat varieties grown at four nitrogen levels.

To limit N-fertilizer applied on wheat, cultivars that use N more efficiently are needed. Our objective was to investigate differences of nitrogen utilization in varieties by studying qualitative and quantitative proteins expression. Two wheat varieties, 'Arche' and 'Récital', were grown under controlled conditions at four N levels (0, 2, 8, and 20 mg N/plant/day) with two replicates. The number of tillers/plant, aerial dry weight/plant and total N content were measured after two months. Two-dimensional gel electrophoresis was also performed on leaf protein extracts. Analyses of variance showed that the N level effect was highly significant for the number of tillers/plant, aerial dry weight and N content. The variety x N level interaction was significant for N content. Analyses of variance on % volume carried out for 524 spots showed a significant variety effect for 55 spots and a significant N treatment effect for 76 spots. Twenty spots showed a significant variety x N treatment interaction. Fourteen proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The possible role of these proteins, eight of which belong to the carbon metabolism, is discussed.